Method for Diagnosing or Determining the Prognosis of Colorectal Cancer (CRC) Using Novel Autoantigens: Gene Expression Guided Autoantigen Discovery
a colorectal cancer and autoantigen technology, applied in the field of colorectal cancer prognosis and autoantigen discovery, can solve the problems of greater success in treating, lack of proper screening and early detection, and 37% of crc cases currently being caught early
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example 1
Gene Expression Analysis of MAP4K4 in Colorectal Cancer
[0034]Gene Expression Analysis of Recurrent vs. Non-Recurrent CRC
[0035]The CRC gene expression dataset was exclusively licensed from Ananomouse Corporation (Cambridge, Mass.) and was produced by whole-genome DNA microarray analysis as follows: The tumor tissue was assayed on the industry standard for oligonucleotide microarrays, the Affymetrix (Santa Clara, Calif.) GeneChip® Human Genome U133 Plus 2.0 array. Analytics were performed utilizing Praxis™ (Ananomouse Corporation, Cambridge, Mass.), a bioinformatics analysis software tool. A component of Praxis™ implements stringent quality assurance metrics to ensure only the highest-quality arrays continue on to the final analysis, which reduces intra-class variability and maintains high signal-to-noise ratios. A power analysis based on preliminary data determined 25 samples were required per class (recurrent and non-recurrent CRC) to achieve greater than 90% power to detect a true ...
example 2
Gene Expression-Guided Proteome Microarray Based Analysis and Discovery of the Novel Colorectal Cancer (CRC) Autoantigen MAP4K4
[0039]The human MAP4K4 novel TAA was analyzed using a high density protein microarrays, to detect autoantibodies in the sera of CRC patients as well as healthy (normal) and autoimmune disease patient controls. As discussed further in the Results section of this Example, this discovery of MAP4K4 as a TAA was guided and facilitated by prior gene expression analysis (see Example 1). It should also be noted that while MAP4K4 was not previously known as a TAA, the mitogen activated protein kinase (MAPK) cell-signaling pathway, and more specifically, MAP4K4 gene expression, have also been associated with CRC in the scientific literature [Hao, Chen, Sui, Si-Ma, Li, Liu, Li, Ding and Li (2010) J Pathol 220: 475-89; Lascorz, Forsti et al. (2010) Carcinogenesis 31: 1612-9].
Serum Screening on Microarrays
[0040]Patient sera were screened against commercial human proteome...
example 3
Validation of Novel Colorectal Cancer (CRC) Autoantigen MAP4K4 Using an ELISA
[0048]The human MAP4K4 TAA was validated using an Enzyme-Linked Immunosorbent Assay (ELISA) to detect autoantibodies in the sera of CRC and healthy (normal) patients.
Enzyme-Linked Immunosorbent Assay (ELISA) of Autoantigen
[0049]Note that some of the CRC and normal patient sera used in the ELISA were the same as used on the ProtoArray® microarrays, while others were not. CRC and normal sera were from Asterand Inc. (Detroit, Mich.), ProMedDx, LLC (Norton, Mass.) and the Ontario Institute of Cancer Research (OICR). A total of 47 normal and 47 CRC sera were used.
[0050]CRC sera were an approximate 50:50 distribution of a) stage T2 or T3 (AJCC staging) non-metastatic and b) stage T3 or T4 metastatic.
[0051]Human MAP4K4 recombinant protein expressed in insect cells and purified by its N-terminal GST fusion tag was purchased from Invitrogen (Carlsbad, Calif.; catalog number PV3687). 384-well white opaque, flat botto...
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